{{'Search' | translate}}
 

Monoclonal Anti-Cytokeratin Peptide 18 antibody produced in mouse

氯化钙二水合物

Company: Sigma-Aldrich
Catalog#: C3306
Bio-protocol()
Company-protocol()
Other protocol()

Preparation of Cerebellum Granule Neurons from Mouse or Rat Pups and Evaluation of Clostridial Neurotoxin Activity and Their Inhibitors by Western Blot and Immunohistochemistry
Author:
Date:
2018-07-05
[Abstract]  Cerebellar Granule Neurons (CGN) from post-natal rodents have been widely used as a model to study neuronal development, physiology and pathology. CGN cultured in vitro maintain the same features displayed in vivo by mature cerebellar granule cells, including the development of a dense neuritic network, neuronal activity, neurotransmitter release and the expression of neuronal protein markers. Moreover, CGN represent a convenient model for the study of Clostridial Neurotoxins (CNT), most notably known as Tetanus and Botulinum neurotoxins, as they abundantly express both CNT receptors and intraneuronal substrates, i.e., Soluble N-ethylmaleimide-sensitive factor activating protein receptors (SNARE proteins). Here, we describe a protocol for obtaining a highly pure ... [摘要]  来自产后啮齿动物的小脑颗粒神经元(CGN)已被广泛用作研究神经元发育,生理学和病理学的模型。 CGN体外培养维持成熟小脑颗粒细胞在体内显示的相同特征,包括发育致密的神经炎网络,神经元活动,神经递质释放和神经元的表达 蛋白质标记。 此外,CGN代表了梭菌神经毒素(CNT)研究的便利模型,最着名的是破伤风和肉毒杆菌神经毒素,因为它们大量表达CNT受体和神经元内基质, ie ,可溶性N-乙基马来酰亚胺 - 敏感因子激活蛋白受体(SNARE蛋白)。 在这里,我们描述了从出生后大鼠/小鼠获得高纯度CGN培养物的方案和用CNT中毒的简便方法。 我们还说明了评估CNT活性及其抑制的方便方法。

【背景】梭菌神经毒素(CNT)的大家族由破伤风神经毒素(TeNT)和肉毒杆菌神经毒素(BoNT)的多种变体形成,它们分别是破伤风和肉毒中毒的神经麻痹毒素(Schiavo et al。,2000; Johnson和Montecucco,2008; Rossetto et al。,2014)。 TeNT,七种BoNT血清型(BoNT / A至/ G)及其许多亚型是金属蛋白酶,通过切割SNARE蛋白(可溶性N-乙基马来酰亚胺敏感因子激活蛋白受体),三种必需蛋白质来阻断神经递质的释放而引起神经麻痹。控制突触小泡与突触前质膜的融合(Rossetto et al。,2014; ...

Hyaluronan Isolation from Mouse Mammary Gland
Author:
Date:
2018-06-05
[Abstract]  The glycosaminoglycan hyaluronan (HA) is a key component of the extracellular matrix. The molecular weight of HA is heterogeneous and can reach from several million to several hundred daltons. The effect of HA on cell behavior is size dependent; fragmented HA acts as a danger signal, stimulates cell migration and proliferation and is proinflammatory, native high molecular weight HA suppresses inflammation. Therefore, it is important to analyze HA size distribution when studying the role of HA in tissue homeostasis and pathology. This protocol describes isolation of HA from mouse mammary glands but can also be applied to other tissues. The quality of the isolated HA is sufficient to analyze size distribution by gel electrophoresis (Calabro et al., 2000). [摘要]  糖胺聚糖透明质酸(HA)是细胞外基质的关键组分。 HA的分子量是不均匀的,可以达到几百万至几百道尔顿。 HA对细胞行为的影响与尺寸有关; 片段化的HA作为危险信号起作用,刺激细胞迁移和增殖并且是促炎性的,原生高分子量HA抑制炎症。 因此,在研究HA在组织稳态和病理学中的作用时,分析HA的大小分布非常重要。 该协议描述从小鼠乳腺分离HA,但也可以应用于其他组织。 分离的HA的质量足以通过凝胶电泳分析大小分布(Calabro等人,2000)。

【背景】糖胺聚糖HA由N-乙酰葡糖胺和β葡萄糖醛酸二糖组成,并且是细胞外基质的普遍存在的组分。 高分子量HA通过酶促降解和被活性氧和氮物质氧化而碎裂。 在健康组织中,大部分HA具有高分子量。 片段化HA的积累在病理过程中起着危险信号的作用(Tolg等人,2012和2017; Yuan等人,2015)。 例如,HA片段刺激炎症,而高分子量HA抑制炎症。 HA通过与细胞膜受体相互作用影响细胞行为,如细胞迁移和增殖,导致信号通路的激活。 由于受体-HA相互作用受HA大小的影响,HA对组织生物学的影响不仅取决于HA量,而且取决于各个细胞的HA大小分布和HA受体表达。

Dual-probe RNA FRET-FISH in Yeast
Author:
Date:
2018-06-05
[Abstract]  mRNA Fluorescence In Situ Hybridization (FISH) is a technique commonly used to profile the distribution of transcripts in cells. When combined with the common single molecule technique Fluorescence Resonance Energy Transfer (FRET), FISH can also be used to profile the co-expression of nearby sequences in the transcript to measure processes such as alternate initiation or splicing variation of the transcript. Unlike in a conventional FISH method using multiple probes to target a single transcript, FRET is limited to the use of two probes labeled with matched dyes and requires the use of sensitized emission. Any widefield microscope capable of sensitive single molecule detection of Cy3 and Cy5 should be able to measure FRET in yeast cells. Alternatively, a FRET-FISH method can be ... [摘要]  mRNA荧光原位杂交(FISH)是一种常用于分析细胞中转录物分布的技术。 当与常见的单分子技术荧光共振能量转移(FRET)相结合时,FISH也可用于分析转录本中附近序列的共表达以测量转录本的替代启动或剪接变异等过程。 与使用多个探针靶向单个转录物的常规FISH方法不同,FRET限于使用用匹配染料标记的两个探针,并且需要使用敏化发射。 任何能够灵敏地检测Cy3和Cy5单分子的宽视场显微镜应该能够测量酵母细胞中的FRET。 或者,可以使用FRET-FISH方法明确确定转录本的身份,而不使用其他FISH技术中使用的引导探针组。

【背景】单细胞转录物分布的定量通常通过用多个探针靶向mRNA来实现,以实现可以与非特异性结合的探针区分开的明亮信号(Raj和Tyagi,2010)。但是,在某些情况下,转录本上有特征,例如剪接变体或替代起始位点,这与常规FISH探针组无法区分。这些同种型序列可以具有短的50nt唯一识别序列。使用两种探针,可以使用FRET对定位结合的任一侧,同时定量多达三种mRNA同种型,例如,具有两种探针的同种型(FRET),具有探针1的同种型仅限于探针2的同种型。依赖于单个荧光团或荧光团对需要通过EMCCD进行灵敏检测。而且,可以使用FRET对(Wadsworth等人,2017)来估计没有其他同种型的序列的探针的检测效率。

Comments